Objective To investigate the feasibility, accuracy and safety of percutaneous pedicle screws placement assisted by navigation with 3D printing template in transforaminal lumbar interbody fusion(TLIF) for lumbar spinal stenosis. Methods From January 2016 to March 2017, the clinical data of single segment lumbar spinal stenosis patients treated with TILF were retrospectively analyzed. Screws placement of 10 patients assisted by 3D printing template navigation were selected as the experimental group, and 10 cases with similar general conditions were selected as the control group. In the experimental group, screws placement on the side with no clinical manifestation assisted by navigation with 3D printing template(template side), and on the other side through Wiltse approach(puncture side). In the control group, pedicle screws were placed by traditional bare-handed method. The operation time, blood loss, fluoroscopy time and frequency, accuracy of pedicle screw implantation and complications of spinal cord and nerve injuries were recorded. Results The operation time and blood loss in the experimental group were less than those in the control group, and the differences were statistically significant(P<0.05). The fluoroscopy time and frequency in template side were less than those in puncture side and the control group, and the differences were statistically significant(P<0.05); while there was no significant difference between the puncture side and the control group(P>0.05). There were no grade 2 screws in the experimental group. There were 4 grade 2 screws in the control group. The percentage of grade 0 pedicle screw, 2 sides of the experimental group were higher than that of the control group, and the differences were statistically significant(P<0.05); while there was no significant difference between the 2 sides of the experimental group (P>0.05). There was no such complications as spinal cord and nerve injuries in 2 groups. Conclusion Percutaneous pedicle screw placement assisted by navigation with 3D printing template in TLIF for lumbar spinal stenosis is feasible. Although it is necessary to avoid the influence of skin movements and postural changes on accuracy of implant nails, it can be regarded as a new and optional technique for minimally invasive nailing.